Compositions and methods for joining non-conjoined lumens

ABSTRACT

Disclosed are compositions, methods, and kits for joining together non-conjoined lumens in a patient&#39;s body including vascular lumens. More particularly, in various aspects, this invention provides compositions, methods, and kits for joining such non-conjoined lumens, including small lumens typically requiring microsurgical technique

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. § 119(e) ofprovisional Patent Application Ser. No. 61/015,660, filed on Dec. 20,2007, which is incorporated herein by reference in its entirety.

FIELD OF THE INVENTION

This invention relates generally to compositions, methods, and kits forjoining together non-conjoined lumens in a patient's body includingvascular lumens. More particularly, in various aspects, this inventionprovides compositions, methods, and kits for joining such non-conjoinedlumens, including small lumens typically requiring microsurgicaltechniques.

BACKGROUND OF THE INVENTION

Non-conjoined lumens arise in a variety of settings including surgicalsettings where the non-conjoined lumens are intentionally created orarise from lacerations or puncture wounds. Intentionally creatednon-conjoined lumens include those arising during surgical repair ofe.g., treatment of a blockage in a lumen by bypass procedures, attachinga synthetic graft or during free tissue transfer in cosmetic surgicalsettings. Anastomosis is conducted to surgically reconnect the open endsof the lumen. Examples of anastomosis procedures include anastomoticprocedures on the vasculature, the vas deferens, the fallopian tubes,the urinary tract, tear ducts, bowel, mammary glands, alimentary ducts,pancreatic ducts, bile ducts, etc. In each case, the anastomosisprocedure creates a channel for the flow of a body fluid there through.

The anastomosis may be, for example, end-to-end, end-to-side, andside-to-side. As is apparent from their names, anastomosis may involvevarious configurations. For instance, one tubular tissue may be joinedend-to-side with two tubular tissues, creating a three-channeled tubulartissue construct.

In the surgical context, end-to-end anastomosis, as is apparent from itsname, is a surgical procedure for connecting an end or distal portion ofone tubular tissue structure to an end or distal portion of anothertubular tissue structure, such that a continuous lumen is created. (Asused herein, “end” or “distal portion,” refers to the open end of thetubular tissue.)

In an end-to-side anastomosis, a tubular tissue structure having a holeor open part is connected through the open part to an open or distal endof a tubular tissue to form a continuous lumen with a branchedconfiguration. Similarly, in a side-to-side anastomosis, twonon-conjoined lumens are merged together into a continuous lumen thougha hole or opening on each of the lumens to be joined.

A successful anastomosis typically involves the smooth connection oflumens, such that the internal structure is not blocked and internalbody fluid flow—such as blood, semen or food or gastrointestinalfluids—is restored or improved. Ideally, the matching up/ligationsurgical procedure is rapid and precise, so that patient exposure whilein a vulnerable state—such as having blood flow stopped—is minimized.

There are a variety of tubular tissues, and the lumen of the firsttubular tissue may not be of the same diameter as the lumen of thesecond tubular tissue. Thus, because the delicate surgery may involvematching and ligating two (or more) non-identical tubular tissues,various ligation techniques have been used with varying rates ofsuccess. These include sutures, tissue adhesives, adhesive strips, andstaples, clips and other devices. To some extent all of these materialsinvolve the skill of the practitioner in anastomosis which is accurate,durable and free from conditions which could cause latent deleteriousreactions in vivo.

The labor-intensive needle and thread remains the most-used technologyas of the present day. Because of the complexity and judgment requiredin suturing, automated techniques are not well accepted. Calcified anddiseased vessels provide mechanical challenges. Sutures may, in someinstances, cause a reaction resulting in long term stenosis or fibrosis.

Other approaches to anastomosis include the use of sealants and biogluesfor ligation. These may be used individually or in conjunction withsuturing or other mechanical ligation techniques or devices. Forexample, one commercially available sealant CoSeal® (AngiotechPharmaceuticals, Inc., Vancouver, B.C., Canada) may complement suturingin cardiovascular surgeries.

Mechanical anastomosis devices, such as clips, are also available. Onecommercially available device, the U-Clip™ (Medtronic, Minneapolis,Minn. 55432 USA), essentially provides a sharp, nitinol hook forknotting to compete anastomosis. The nitinol allows reversibledeformation. The C-Port® (Cardica, Inc. Redwood City, Calif. 94063 USA)and related products are commercially available and use miniaturestainless steel staples to securely attach the bypass graft to thecoronary artery.

But, before ligating end-to-end, for example, the practitioner mustmatch up the lumens by the circumference of the vessel, using bloodvessels as an illustration. Frequently, this is troublesome to thepractitioner because the end of an tubular tissue—such as a clampedblood vessel devoid of blood—is not a perfectly round circle; rather itis in its unpressurized, deflated-looking state where a cross-sectionalview of the circumference may be a circle, an oval or irregular, and, ofcourse having no structural support from within, is unstably in anyshape (unless the surrounding tissue possesses structural strength). Thesize of the vessels to be so connected may be different. Although bloodvessels, for example, or other tubular tissues are somewhat elastic(deformable and returning to the original shape) or plastic (deforming,and not fully returning to the original shape), connecting thecircumferences of the lumens such that upon ligation there is no orminimal leakage (in the vascular context, for example), requires askilled practitioner.

In a microvascular context, anastomosis is performed between ends ofblood vessels in the course of, for example, reattaching severed bodyparts or transplanting organs or tissue. Microvascular anastomosis isoften performed by hand under a microscope, and is tedious andpainstaking work. The blood vessels connected together often havedifferent diameters, both of which are very small, on the order of about1 to about 5 millimeters (“mm”). Although blood vessels are usually atleast somewhat elastic, the practitioner must match up end to end (forexample) two different-shaped-different-sized circumferences and thenstitch them together (for example). As a result, it can take many hoursto complete just the microvascular anastomosis required to reconnect asevered body part or transplant an organ.

One attempt to provide a mechanism for performing such a microvascularanastomosis is the Microvascular Anastomotic Coupler System, availablefrom Bio-Vascular, Inc. (San Diego, Calif., USA). In this mechanism, anend of each vessel to be connected is essentially turned outward(“everted”) over a ring with a forceps or similar instrument. Each ringincludes a number of pins over which the vessel is everted. The ringsare then pressed together, such that the pins on each ring enterrecesses in the other ring, connecting the rings and holding the ends ofthe vessels together. This system, however, is limited to use with twoblood vessels having substantially the same diameter. Further, manualeversion of a blood vessel having a diameter on the order of onemillimeter is difficult and painstaking, particularly when the eversionis to be substantially even around the circumference of the ring.Further, the rings provide a noncompliant anastomosis between the twovessels. Thus, although stabilizing the circumference facilitates theability of the practitioner to match up vessels for end-to-endmicrovascular anastomosis, the device requires, essentially,practitioners skilled in microsurgical techniques.

For patients and practitioners, perhaps the most demanding anastomosisis incident to heart revascularization. The arteries that bring blood tothe heart muscle (coronary arteries) can become clogged by plaque (abuildup of fat, cholesterol and other substances). This can slow or stopblood flow through the heart's blood vessels, leading to chest pain or aheart attack. Increasing blood flow to the heart muscle can relievechest pain and reduce the risk of heart attack. A patient may undergoone, two, three or more bypass grafts, depending on how many coronaryarteries are blocked.

Coronary artery bypass graft surgery (“CABG”, sometimes pronounced“cabbage” by practitioners) reroutes, or “bypasses,” blood aroundclogged arteries to improve blood flow and oxygen to the heart. Inperforming the CABG anastomosis, a segment of a healthy blood vesselfrom another part of the body is used to make a detour around theblocked part of the coronary artery. This healthy blood vessel may be,for example, an artery present in the thoracic cavity, or may be a pieceof a long vein from the patient's leg. In some circumstances, graftsfrom non-autologous sources may be used, such as synthetic tubulartissues or animal tubular tissues. Regardless of the source of thehealthy blood vessel, one end is connected to the large artery leavingthe patient's heart (the aorta), and the other end is attached or“grafted” to the coronary artery below the blocked area. In this way of“rewiring” the vasculature, substantially unobstructed blood flow to theheart muscle is resumed.

Conventionally, a pump oxygenator (heart-lung machine) is used forcoronary bypass graft operations. Medicines are used to stop thepatient's heart, which allows the practitioner to operate without theheart beating. The heart-lung machine keeps oxygen-rich blood movingthroughout the patient's body. For this conventional heart bypass graftsurgery, a team of practitioners is needed (a surgeon, cardiacanesthesiologist and surgical nurse, and a perfusionist (blood flowspecialist)). Multiple practitioners, additional complexity, and, as apractical matter, additional health care cost is involved over surgicalprocedures involving fewer practitioners and procedures.

Moreover, blood quality may be degraded as the heart-lung machinerepetitively pumps the patient's blood through the systemic circulation.The blood may embolize or clot in the distal circulation, or form clotswhich migrate to the distal vasculature, and cause a stroke.

Off pump coronary artery bypass surgery may reduce this risk. “Off Pump”coronary artery bypass grafting, also called beating heart bypassgrafting, takes place while the heart continues to beat, but amechanical device may be used in an attempt to steady the surroundingvasculature, so that the practitioner can perform the graft. Frequently,because the graft must be performed on arteries in locations directlyaffected by the beating heart, stabilizing mechanisms are not thoroughlyeffective, and the practitioner must suture the graft while the graft ismoving in conjunction with the heart beat, at least to some extent.Thus, the graft quality may be compromised.

Although, in a bypass surgery time is of the essence, the practitionercannot rush through without thoroughly and precisely anastomising thegraft(s). In conventional coronary artery bypass surgery, three criticaldeterminates that affect the outcome of a bypass surgery are:

-   -   (1) time the patient spends on cardiopulmonary bypass,    -   (2) time the patient spends with a clamped aorta, and    -   (3) the quality of the anastomosis.

After an hour, the risk of patient morbidity is thought to increaseperhaps due to the heart-lung machine degrading the quality of the bloodas it is circulated through the systemic circulation. Bypass surgeries,however, often last three hours or longer. Moreover, where the aorta isclamped and blood therefore cannot pass through, the blocked blood isthought to cause additional issues.

Anastomosis is time-consuming. The average time for suturing oneanastomosis is approximately fifteen to sixty minutes. An average CABGprocedure is thought to involve approximately five anastomoses.Therefore, the average time for graft suturing exceeds the sixty-minutethreshold for increased patient morbidity. Patients treated withconventional coronary surgery and placed on cardiopulmonary bypass wouldbenefit from reducing the amount of time spent performing eachanastomosis.

In “off pump” procedures where the heart remains beating, the difficultyof suturing an anastomosis graft on a moving surface of the heart maydegrade the quality of such grafts completed on patients. An anastomosisdiffers from straight line suturing in that each suture has a differentorientation that is based on its position around the cross-sectionalcircumference of the blood vessel graft. It can be appreciated that someof the sutures are easily made from on top of the conduit or bloodvessel graft, while others are more difficult to complete as they arebeneath the conduit. It can be further appreciated that performing suchcomplex suturing procedures on a moving platform, such as the beatingheart, further increases the difficulty associated with such suturingprocedures. Improperly connecting blood vessel grafts to the patient maypresent substantial post-operative complications and/or increaseoperating room time spent correcting the improperly connected graft.

Accordingly, for surgical anastomosis, both practitioners and patientswould benefit from faster procedures allowing patients to minimizeprocedure time, and simpler methods allowing reduced complexity, ease ofuse and higher quality anastomosis with fewer complications.

SUMMARY OF THE INVENTION

This invention is directed to the discovery that the stabilization ofthe geometry of the distal portions of tubular tissues or lumensfacilitates joining such non-conjoined lumens.

In one aspect, this invention provides novel thermoreversible sol-gelcompositions for surgical use to stabilize the geometry of the terminalportion of a lumen to be joined with another lumen, which compositionshave a phase transition temperature from a liquid phase to a gel phaseof at least 1° C. above or below the temperature of the surgical fieldand an elastic modulus of at least 8,000 Pascals. In some embodiments,the compositions comprise at least one poloxamer in an aqueous solvent.In some embodiments, the compositions comprise at least two poloxamersin an aqueous solvent.

In some embodiments, the invention provides a composition whichcomprises:

-   -   a) about 25 to 33% of a mixture of poloxamer 407 and poloxamer        188 in a ratio of between 3:1 and 0.8:1;    -   b) an aqueous salt solution having an ionic strength of 0.05 M        to 0.4 M;    -   c) a phase transition temperature from a liquid phase to a gel        phase of at least 1° C. above or below the temperature of the        surgical field; and    -   d) an elastic modulus of at least 8,000 Pascals in the gel        phase.

In another aspect, this invention provides a method of joining at leasttwo non-conjoined lumens in a patient, which method comprises:

-   -   a) applying a thermoreversible sol-gel composition in at least        the distal portion of at least one of the lumens in a manner        which imparts structural integrity to said portion of the lumen        or lumens, which composition has a phase transition temperature        from a liquid phase to a gel phase of at least 1° C. above or        below the temperature of the surgical field and an elastic        modulus of at least 8,000 Pascals in the gel phase, which        composition comprises at least two poloxamers in an aqueous        solution;    -   b) aligning the lumens;    -   c) joining the aligned lumens to form a conduit; and    -   d) removing the composition thereby establishing body fluid flow        through the conduit.

In one embodiment, the method comprises:

-   -   a) applying in a gel phase a thermoreversible sol-gel        composition into at least the distal portion of at least one of        the lumens which composition has a phase transition temperature        from a liquid phase to a gel phase of at least 1° C. above or        below the temperature of the surgical field and an elastic        modulus of at least 8,000 Pascals in the gel phase, which        composition comprises at least two poloxamers in an aqueous        solvent;    -   b) aligning the lumens;    -   c) joining the aligned lumens to form a conduit; and    -   d) removing the composition thereby establishing body fluid flow        through the conduit.

In another embodiment, the method comprises:

-   -   a) applying in a solution phase a thermoreversible sol-gel        composition into at least the distal portion of at least one of        the lumens which composition has a phase transition temperature        from a liquid phase to a gel phase of at least 1° C. above or        below the temperature of the surgical field and an elastic        modulus of at least 8,000 Pascals in the gel phase, which        composition comprises at least two poloxamers in an aqueous        solvent;    -   b) transitioning the sol-gel composition from a liquid phase to        its solid phase;    -   c) aligning the lumens;    -   d) joining the aligned lumens to form a conduit; and    -   e) removing the composition thereby establishing body fluid flow        through the conduit.

In another aspect, this invention provides methods for imaging thejoinder of at least two non-conjoined lumens using the methods of thisinvention and the visualization of the geometry of the distal portionsof the lumens in a patient using a sol-gel composition of thisinvention.

In another aspect, this invention provides methods for visualizingguidance for the methods of this invention for joining at least twonon-conjoined lumens in a patient using a sol-gel composition of thisinvention.

In another aspect, this invention provides a method for evaluating theeffectiveness of the methods of this invention for joining non-conjoinedlumens in a patient using a sol-gel composition of this invention.

In another aspect, this invention also includes use of any of thematerials or methods as disclosed herein for manufacture of a medicamentfor joining lumens, particularly in a live patient, as further describedherein. Thus, in one embodiment, the present invention provides abiocompatible solid mass for use in joining at least two non-conjoinedlumens in a patient in a method which comprises:

-   -   a) placing the a sol-gel composition of this invention in at        least the distal portion of at least one of the lumens;    -   b) aligning the lumens;    -   c) closing the aligned lumens to form a conduit; and    -   d) removing the solid mass thereby establishing flow through the        conduit.

In still another aspect, this invention provides novel kits useful inthe methods described above.

These and other embodiments of this invention are further described inthe text below. The methods, compositions and kits of this invention maybe used in both human and non-human mammals.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention is best understood from the following detailed descriptionwhen read in conjunction with the accompanying drawings. It isemphasized that, according to common practice, the various features ofthe drawings are not to-scale. On the contrary, the dimensions of thevarious features are arbitrarily expanded or reduced for clarity.Included in the drawings are the following figures:

FIG. 1 is a schematic cross-sectional view of a tubular tissue with apartial blockage.

FIG. 2 is a schematic cross-sectional view of the tubular tissue of FIG.1 wherein the area having the partial blockage has been removed creatingtwo non-conjoined lumens having both ends of the tubular tissue clampedoff.

FIG. 3 is a schematic cross-sectional view of the tubular tissue of FIG.1 with one end filled with thermoreversible sol-gel (solid or gel phase)and the other end being filled with the sol-gel (liquid phase) via asyringe.

FIG. 4 is a schematic cross-sectional view of the tubular tissue of FIG.1 with the ends sealed, the gel (solid phase) in place and the clampsstill present.

FIG. 5A is a schematic cross-sectional view of the tubular tissue ofFIG. 1 with the ends sealed, the sol-gel (reversed to liquid phase) inplace and the clamps still present.

FIG. 5 is a schematic cross-sectional view of the tubular tissue of FIG.1 with the ends sealed, the sol-gel (reversed to liquid phase) in placeand the clamps removed.

FIG. 6 is a cross-sectional view of the tubular tissue of FIG. 1 showingthe sol-gel dissolved and flow restored with the blockage area removed.

DETAILED DESCRIPTION OF THE INVENTION

Before the present compositions, medical systems, kits, and methods aredescribed, it is to be understood that this invention is not limited toparticular embodiments described, as such may, of course, vary. It isalso to be understood that the terminology used herein is for thepurpose of describing particular embodiments only, and is not intendedto be limiting, since the scope of the present invention will be limitedonly by the appended claims.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise. Thus, for example, reference to “athermoreversible gel” includes a plurality of such gels and reference to“the adhesive” includes reference to one or more adhesives andequivalents thereof known to those skilled in the art, and so forth.

I. DEFINITIONS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. As used herein the followingterms have the following meanings. If not defined, a term has its artrecognized meaning.

The term “about” when used before a numerical designation, e.g., pH,temperature, amount, concentration, including range, indicatesapproximations which may vary by (+) or (−) 10%, 5%, 1% or 0.1%.

As used herein, the term “comprising” or “comprises” is intended to meanthat the compositions and methods include the recited elements, but notexcluding others. “Consisting essentially of” when used to definecompositions and methods, shall mean excluding other elements of anyessential significance to the combination for the stated purpose. Thus,a composition consisting essentially of the elements as defined hereinwould not exclude other materials or steps that do not materially affectthe basic and novel characteristic(s) of the claimed invention.“Consisting of” shall mean excluding more than trace elements of otheringredients and substantial method steps. Embodiments defined by each ofthese transition terms are within the scope of this invention.

The term “patient” refers to mammals and includes humans and non-humanmammals.

The term “surgical” relates to any medical procedure where the innerorgan or issue of a patient is accessed to investigate and/or treat apathological condition such as disease or injury, to help improve oralter bodily function or appearance, or for other reasons. As usedherein, “surgical” relates to procedures of accessing a patient's innerorgan or tissue via an incision or an opening on the patient and vianeedle-puncture of the skin, such as in percutaneous approaches andother minimally invasive procedures, such as laparoscopic surgeries.

The term “lumen” refers to the hollow tube and the surrounding tissuedefining the hollow tube, such as a blood vessel, a vas deferens, afallopian tube, urinary tract, a tear duct, bowel, a mammary gland, analimentary duct, a pancreatic duct, a bile duct, and the like. Lumensalso includes artificial conduits, such as ePTFE grafts. The term“lumen” is used interchangeably with the term “duct.”

The term “conduit” refers to the hollow tube formed by the joined lumenswhich can accommodate flow of a body fluid after removal of the sol-gelplaced therein or clamps placed thereon.

The term “providing” as used in, for example, “providing a sol-gelcomposition in a distal end of a lumen” and the like, refers to the actof causing a sol-gel composition to be present in the lumen to supportthe structural integrity of the lumen. The act can be “placing”,“delivering” or the like, the sol-gel can be placed in the lumen in asolid state or in a liquid state and being transferred into a solidstate while inside the lumen.

The term “non-conjoined lumen” refers to a lumen having an open end or ahole where a body fluid that the lumen carries or should carry willundesirably leak out if the open end or hole is not occluded orconnected to another lumen to form a continuous lumen in which the bodyfluid can circulate or be delivered to the desired destination withoutundesired leakage.

The term “aligning” refers to the act of bringing the lumens to bejoined in a position that the lumens may be connect in a desirablemanner, which may include matching up the ends of the lumens and pushingthe matched ends of the lumens in contact of each other. Preferably,aligning is done without causing substantial tension to the lumens andto the respective tubular tissues they connect to.

The term “approximating” refers to the act of bringing the lumens to aposition or to a close proximity of a position that they are to beconnected. When two lumens are approximated, they are close to eachother but there may be gaps between the ends of the lumens so that asol-gel composition can be applied to the inside of the lumens toprovide structural support.

The term “biocompatible” as used with terms such as “biocompatiblepolymer”, “biocompatible compound” and the like, refer to materialswhich, in the amounts employed, are non-toxic and substantiallynon-immunogenic when used internally in a subject such as a humanpatient.

The term “thermoreversible sol-gel” refers to a composition, whichundergoes a phase transition from a liquid phase to gel phase when thetemperature is raised above or reduced below a critical value, which isreferred to as “transition temperature.” The term “liquid phase” or“liquid state” refers to a liquid or flowable form with a viscosity ofless than 2000 Pascal-seconds. The term “gel phase” or “gel state”refers to a gel or relatively solid form with a viscosity of greaterthan 10,000 Pascal-seconds. As is well known in the science of rheology,complex viscosities of compositions are reported in Pascal-seconds andall viscosities reported in this application are reported as complexviscosities. Such phase transition is reversible. Thus, athermoreversible sol-gel composition changes from its liquid state toits gel state when the temperature is raised to or above the criticalvalue, or transition temperature, and undergoes a phase transition fromthe gel state to the liquid state when the temperature is lowered to orbelow the critical value, or transition temperature. Preferably thephase transition from a liquid to a gel and vice versa occurs in lessthan 10 minutes, more preferably in less than 5 minutes and even morepreferably in less than 2 minute.

For the purpose of this invention, it is desired that the gel hassufficient stiffness or “elastic modulus” to maintain the shape of thefilled lumen (i.e. impart structure integrity) during joining of thedistal opening of the lumens. The term “gel phase elastic modulus”refers to the stiffness of the sol-gel composition when it is in its gelstate or at a temperature above the transition temperature. The term“liquid phase elastic modulus” refers to the stiffness of the sol-gelcomposition when it is in its liquid state or at a temperature below thetransition temperature.

In one embodiment, the thermoreversible sol-gel composition has amodulus of from about 8,000 to 50,000 or 10,000 to 50,000 Pascals and,more preferably, from about 12,000 to 40,000 Pascals.

The term “aqueous solvent” refers to water or a water based solution,e.g. an aqueous salt solution, such as a saline solution, phosphatebuffered saline, and other aqueous solutions suitable for dissolving thepoloxamers described herein. An aqueous salt solution may contain one ormore biocompatible salts selected from sodium chloride (NaCl), potassiumchloride (KCl), sodium sulfate (Na₂SO₄), sodium bisulfate (NaHSO₄),sodium phosphate (Na₃PO₄), monosodium phosphate (NaH₂PO₄), disodiumphosphate (Na₂HPO₄), potassium phosphate (K₃PO₄), monopotassiumphosphate (KH₂PO₄), dipotassium phosphate (K₂HPO₄), various solublecalcium and magnesium salts, such as calcium chloride (CaCl₂), magnesiumchloride (MgCl₂) and other salts formed by a combination of a cationselected from the group consisting of sodium, potassium, calcium,magnesium, ammonium, and tetraalkylammonium, with an anion selected fromthe group consisting of chloride, bromide, tartrate, mesylate, acetate,maleate, and oxalate and other biocompatible, water soluble saltsincluding those described in P. Heinrich Stahl, Camille G. Wermuth(Eds.), Handbook of Pharmaceutical Salts Properties, Selection, and Use;2002.

The term “phosphate buffered saline” or “PBS” refers to a buffersolution which help to maintain a physiological pH and a physiologicalionic strength (i.e. isotonic). The final salt concentration of PBS isapproximately 137 mM sodium chloride (NaCl), 10 mM phosphate, 2.7 mMpotassium chloride, with a pH of 7.4. It may be prepared by diluting astock solution ten times, which stock solution is prepared by dissolving800 g sodium chloride (KCl), 20 g potassium chloride, 144 g disodiumphosphate (Na₂HPO₄) and 24 g monopotassium phosphate (KH₂PO₄) in 8 L ofdistilled water, and topping up to 10 L. PBS can have additional ionssuch as Calcium (Ca²⁺) or Magnesium (Mg²⁺).

The term “the temperature of the surgical field” refers to temperatureof the site where the surgical procedure of joining the non-conjoinedlumens is performed. In some embodiments, the temperature of thesurgical field refers to the temperature of the vessel wall. Typically,the temperature of the surgical field is close to the body temperatureof the patient, but may be somewhat lower than the body temperature ofthe patient. The difference between the temperature of the surgicalfield and the patient's body temperature can be affected by a number offactors, such as the size of the incision made in the patient's body,the duration of the exposure, the temperature of the operation room,etc.

The term “distal end” and “opening” of a lumen are used interchangeablyherein and refer to the opening of the lumen, for example, the two endscreated when a lumen is surgically divided into two parts. “Distal end”or “opening” also refers to a hole cut on part of the wall of a lumenalthough the lumen is not completely divided, as in the case of anend-to-side or side-to-side anastomosis.

The term “distal portion of the lumen” refers to the portion of thelumen adjacent to the opening in the lumen. Thus, for example, when alumen is surgically cut, the resulting two openings define the distalportion of what are now first and second lumens. Distal portion of thelumen also refers to the portion adjacent to the hole of a lumen to beused in an end-to-side or side-to-side anastomosis procedure.

The term “joining” refers to any method wherein the first and secondlumens are structurally joined together including by way of example,suturing, use of biocompatible glues, lasers, etc. In a preferredembodiment, joining of the lumens is conducted under conditions wherethere is little or no leakage of body fluid from the juncture of thejoined lumens.

The term “substantial completion” as used in, for example, “confirmingsubstantial completion of phase change,” refers to the amount of thephase change that would ensure that the sol-gel composition can besafely removed from the anastomosis site by the flow of body fluid inthe lumen without causing undesirable blockage in the lumen, for examplein a portion of a blood vessel connected to and is smaller than thevessel being joined. In some embodiments, “substantial completion”refers to equal to or greater than about 50%, 80%, 90%, or 95% and up to100% phase change.

The term “removing the thermoreversible sol-gel composition” refers toremoving the sol-gel composition from the conduit formed by the joinedlumens. This can be done by transitioning the thermoreversible sol-gelcomposition from the gel phase to a liquid phase by lowering thetemperature to below its transition temperature or by dissolutionfollowed by flushing with the body fluid inside the lumen. Further, anycomplementary methods of turning the thermoreversible sol-gelcomposition into a liquid flowable form can also be employed. Forexample, certain thermoreversible sol-gel compositions can undergo phasetransition from a gel to a liquid when triggered by a change in ionicstrength, light, pH, etc, alone or in combination with a temperaturechange. Two or more methods can be combined to facilitate removal of thecomposition. After the thermoreversible sol-gel composition is removedfrom the conduit, it will be engrained in the body fluid and become partof the systemic flow of that fluid until removed or cleared by the body.

A “therapeutically active moiety” will be a biologically active moiety.A therapeutic effect is one which seeks to treat the source or symptomof a disease or physical disorder. The term “treat” or “treatment” asused herein refers to: (i) preventing a disease or disorder fromoccurring in a subject which may be predisposed to the disease orcondition but has not yet been diagnosed as having it; (ii) inhibitingthe disease or disorder, i.e., arresting its development; and/or (iii)ameliorating or relieving the disease or disorder, i.e., causingregression of the disease. A therapeutically effective amount issufficient to establish causation of a therapeutic effect, as determinedby relevant clinical standards. The therapeutically effective amountwill vary depending upon the specific agent incorporated, the subjectand disease condition being treated, the weight and age of the subject,the severity of the disease condition, the dosing regimen to befollowed, timing of administration, the manner of administration and thelike, all of which can be determined readily by one of ordinary skill inthe art.

The term “arresting” the flow of a body fluid refers the act ofstopping, substantially stopping, or reducing the flow of the bodyfluid. This can be done a number of ways, such as by using a clamp,providing a vessel loop, silk tie, applying a direct pressure (forexample, using a finger or tournequette).

The term “clamp” refers to conventional mechanical devices suitable forapplication to a lumen to arrest the flow of body fluid therein, forexample, stopping or reducing the flow of blood in a vessel, where thedevices can be retrieved after they are no longer needed, therebyrestoring flow. Clamps include any surgical clamps suitable for clampingthe vessels to be operated on from the outside, such as clamps, clipsand tourniquets or snares. Clamps also include devices that are appliedto the inside of a lumen, such as a balloon catheter that is placedinside a blood vessel and inflated to stop blood flow. Suitable clampsare known in the art.

The terms “adhesive”, “surgical adhesive”, “glue”, “biocompatible glue”or “sealant” and the like are used interchangeably herein. These termsare used to describe compounds which are or can be used in binding onetissue to another tissue. The glue may operate by the formation ofcovalent bonds and allow the tissues to contact each other and naturallyheal or grow together. The adhesive may be comprised of acyanoacrylate-based adhesive, a fibrin-based adhesive, apolyurethane-based adhesive, a polyisocyanate-based adhesive apolyethylene glycol-based glue, a latex glue, a biologics glue (alsoknown as a protein-based glue, such as those comprising serum albumin(such as human, bovine, or porcine serum albumin), and glutaraldehyde),or an ultraviolet curable glue. The adhesives may include anybiocompatible material which when added to the adhesive, produces anopen cell geometry upon curing in situ to promote tissue ingrowth. Forexample, such material can include a foaming agent or porogen, such assodium chloride, crystals of saccharose, gelatin spheres or paraffinspheres, or an emulsified liquid that is compatible but is immisciblewith the glue and can be absorbed by the tissue relatively quickly toleave pores in the glue. Other suitable biocompatible agents are knownin the art. It is contemplated that the tissue can infiltrate the gluematrix and heal across as the glue is being reabsorbed by the body.Adhesive materials may be found within publications known to thoseskilled in the art and reference made to U.S. Pat. No. 7,044,982 issuedMay 16, 2006 and U.S. Pat. No. 6,939,364 issued Sep. 6, 2005. Both ofwhich are incorporated herein by reference along with the publicationscited therein to disclose and describe surgical adhesives to the extentthat these disclosures do not contradict the present disclosure.Examples of glues also include, but are not limited to, ArterX® (byTenaxis Medical, Inc., Mountain View, Calif.), CovaBond® (by CovalentMedical, Inc., Ann Arbor, Mich.), ProGEL VS™ (Neomend, Irvine, Calif.),PPCA & DermaFlex® (by Chemence Medical, Alpharetta, Ga.), Bioglue®(known to be a mixture of bolvine serum albumin and gluderaldehyde, byCryolife, Inc., Kennesaw, Ga.), Coseal® (by Baxter, Deerfield, Ill.),Microval® (by Medico Corp., Bucuresti, Romania), Omnex® (by EthiconInc., Somerville, N.J.), HistoAcryl Blue® (by Aesculap subsidiary of BBraun, in partnership with TissueSeal), Indermil® (by Syneture.Mansfield, Mass.). LiquiBand® (by Advanced Medical Solutions Group, UK),Glubran® (by Gem s.r.l, Italy), GluSeal® (by GluStitch, Canada),Neuroacryl® and TruFill® (by Cordis, Warren, N.J.), and the like. Gluesmay also include Tissucol® and Tisseel® (by Baxter, Deerfield, Ill.),Avitene® (by Davol, Inc. Providence, R.I.), DuraSeal® (by ConfluentSurgical, Waltham, Mass.), Dermabond®, Evicel® and Quixil® (by EthiconInc., Somerville, N.J.), LTG® (by MediGlue, Russia), Epiglu® (byMeyer-Haake GmbH Medical Innovations, Germany), TachoSil® (by Nycomed,Austria) and TissuGlue® (by Cohera Medical, Inc., Pittsburgh, Pa.).

II. COMPOSITIONS

In one aspect, this invention provides a thermoreversible sol-gelcomposition having unique properties for use in the methods of thisinvention for joining two or more non-conjoined lumens, for exampleblood vessels, in a patient. In an illustrative embodiment of the method(a detailed description of the methods of this invention can be found insection “III. Anastomosis Methods” below), two clamps are placed on thevessels to stop blood flow in the section between the clamps prior tosevering the vessel, as shown by FIG. 2 (which illustrates the vesselafter it has been severed). Thus, it is understandable that due to theoutside pressure from the clamps and from manipulations by a surgeonduring the anastomosis, coupled with lack of inside pressure in theabsence of blood flow, the severed vessels are unlikely to maintaintheir structural integrity, which will cause difficulties in joining thevessels. Thus, this invention provides a thermoreversible sol-gelcomposition that can be conveniently placed in the lumens distal to theclamping to provide structural support to the vessels during the joiningprocedure and can be easily removed after the lumens are joined.

In order to provide sufficient structural support, it is contemplatedthat the gel state of the sol-gel composition needs to have a minimumstiffness or elastic modulus of at least 8,000 Pascals, 10,000 Pascals,or 12,000 Pascals. In addition, in order to be used safely in a patient,the composition is preferably biocompatible, sterile and stable and havea transition temperature that is within the range tolerable by humantissues. It is preferred that the thermoreversible sol-gel compositionhas a pH and ionic strength similar to physiological conditions in orderto avoid damage to surrounding tissues.

It is contemplated that a transition temperature of at least 1° C. aboveor below the ambient temperature of the surgical field is used. However,the transition temperature should not exceed 45° C. or be lower than 0°C. in order to be safe to a patient. In some embodiment, the compositionhas a transition temperature of at least 1° C. above or below theambient temperature of the surgical field so that it can readily undergoa thermal sol-gel transition under the surgical setting, which may beimportant to minimize the duration and improve the quality of theprocedure. In some embodiment, the composition has a transitiontemperature of at least 3° C. above or below the ambient temperature ofthe surgical field.

In some embodiments, the thermoreversible sol-gel composition has atransition temperature of between 33° C. to 38° C., or between 35° C. to37° C. In some embodiment, the transition temperature is about 35° C.Such a sol-gel composition is a liquid when stored at room temperatureand can be delivered to the lumen as a liquid. The composition can thenbe slightly warmed by a heat source to above the transition temperatureto form a gel having a desired stiffness. After the vessels are joined,the heat source can be removed to reduce the temperature to or below theambient temperature of the surgical field, inducing the gel to change toits liquid state, which upon restoration of blood flow can be dilutedand removed from the joined vessel. Compositions with a transitiontemperature that is lower than the ambient temperature of the surgicalfield can be delivered as a gel and removed by lowering the temperatureto a degree that is lower than the transition temperature but is stillwithin the range the lumen tissue can tolerate. In order to be easilyremoved from the joined vessel after the anastomosis procedure, it iscontemplated that the thermoreversible sol-gel composition should havegood aqueous solubility and a liquid phase elastic modulus of no morethan 10,000 Pascals so that it can be readily removed and not causeobstruction to the flow of body fluid, such as blood.

In one embodiment, this invention provides a thermoreversible sol-gelcomposition for providing structural support for a lumen to be joinedwith another lumen during a surgical operation in a patient thatsatisfies the above desired parameters. In one embodiment, the sol-gelcomposition of this invention has a phase transition temperature from aliquid phase to a gel phase of at least 1° C. above or below the ambienttemperature of the surgical field and a gel phase elastic modulus of atleast 8,000 Pascals, 10,000 Pascals, or 12,000 Pascals at a temperatureabove the transition temperature. In one embodiment, the transitiontemperature of the thermoreversible sol-gel composition is at least 3°C. above the ambient temperature of the surgical field. In oneembodiment, the composition comprises at least one poloxamer in anaqueous solvent. The aqueous solvent may be water or a biocompatibleaqueous salt solution. In one embodiment, the composition comprises atleast two poloxamers in an aqueous solvent. In another embodiment, thesol-gel composition has a maximum liquid phase elastic modulus of 10,000Pascals.

In one embodiment, the thermoreversible sol-gel composition has atransition temperature of between 35° C. and 38° C. In one embodiment,the composition is a liquid having a maximum elastic modulus of 10,000Pascals at 34° C. and is a gel having a minimum elastic modulus of12,000 Pascals at 37° C. In one embodiment, the composition has anelastic modulus of at least 17,000 Pascals at 40° C.

In one embodiment, the composition is sterile.

In one embodiment, the pH of the composition is 5-9. In anotherembodiment, the pH of the composition is 6-8.

It is a result of this invention that a mixture of poloxamer 188 andpoloxamer 407, given the right concentration and ratio, generatesthermoreversible sol-gel compositions having the above desiredproperties. Poloxamers are biocompatiblepolyoxyethylene-polyoxypropylene block copolymers represented by theformula of HO(C₂H₄O)_(a)(C₃H₆O)_(b)(C₂H₄O)_(a)H that are also known bytheir tradenames such as Pluronics® and Lutrol®. There are several typesof poloxamers based on the values of a and b in the formula and theirmolecular weight, for example, poloxamers 124, 182, 188, 237, 338 and407. When dissolved in water or an aqueous solvent, they formthermoreversible sol-gel compositions. However, certain mixtures ofpoloxamer 188 and poloxamer 407 are found to be of particular use in themethod of this invention. Poloxamer 188, which is also known asPluronic® F68 or Lutrol® F68, has an average molecular weight of about8,400. Poloxamer 407, which is also known as Pluronic® F127 or Lutrol®F127, has an average molecular weight of about 12,5000. It is to beunderstood that a polymer may contain mixtures of varying molecularweights incidental to the polymerization process.

It is discovered that both the ratio of poloxamer 188 and poloxamer 407and the total poloxamer concentration affect the properties of theresulting thermoreversible sol-gel composition. At the high limit of thetotal poloxamer concentration, solubility is a concern. The solubilitylimit of the total concentration of poloxamers 188 and 407 is about 35%.If the total poloxamer concentration is too low, however, thecomposition will not be sufficiently stiff when solidifies. At a givenratio of poloxamer 188 and poloxamer 407, an increase in total poloxamerconcentration causes an increase in elastic modulus but a decrease inliquid-solid transition temperature. A very low transition temperaturemay cause problems in delivery and removal of the thermoreversiblesol-gel composition because the composition may stay in its gel state atthe ambient temperatures of the surgical field, and thus may requirecooling to change to its liquid state before it is removed, complicatingthe anastomosis procedure.

At a given total poloxamer concentration, the transition temperature canbe increased by increasing the proportion of the lower molecular weightpoloxamer, i.e. poloxamer 188. If the composition is rich in poloxamer188, the transition temperature will be too high and in order to inducea liquid to gel transition, the temperature of the lumen may have to beraised to a temperature that may cause tissue damage, making it unsafefor its intended purpose. If the composition is rich in poloxamer 407,the transition temperature will be too low. Generally, if twothermoreversible sol-gel compositions have the same transitiontemperature, the composition with a higher total poloxamer concentrationwill have a higher gel phase elastic modulus. Thus, to increase theelastic modulus but maintain a desired transition temperature, one needto increase the total concentration of poloxamer and the proportion ofpoloxamer 188 simultaneously, and vice versa.

Table 1 provides several working examples of the thermoreversiblesol-gel composition of this invention as well as examples that do notpossess the desired properties for use in the methods of this invention.As shown in Table 1, Formulations 1-3, possess the desired transitiontemperature and stiffness for use in the methods of this invention. Aconstant transition temperature can be obtained through increasing thetotal poloxamer concentration and the poloxamer 188 ratiosimultaneously. The stiffness increases with an increase in totalpoloxamer concentration. However, unbalanced alterations may lead tocompositions unsatisfactory the methods. For example, a 28% totalconcentration combined with a ratio of poloxamer 407 to 188 of 1.55:1(Formulation 1) gives the desired transition temperature and stiffness.However, increasing the ratio of poloxamer 407 to 2.85:1 (Formulation 5)makes the transition temperature too low so that at 34° C. thecomposition is still a gel. On the other hand, decreasing the ratio ofpoloxamer 407 to 1:1 (Formulation 4) makes the transition temperaturetoo high so that at 40° C., the composition is still a liquid. However,a ratio of poloxamer 407 to 188 of 1:1 is operational if the totalpoloxamer concentration is increased from 28% to 31%.

TABLE 1 Transition temper- Modulus Modulus Modulus Formu- Total 407:188ature at 40° C. at 37° C. at 34° C. lation Conc. Ratio (° C.) (kPa)(kPa) (kPa) 1 28% 1.55:1 35 17.6 13.9 4.4 2 31%   1:1 35 23.5 17.1 6.2 333%  0.8:1 35 39.7 28.8 2.6 4 28%   1:1 >40 0.5 0.4 0.3 5 28% 2.85:1 <3319.8 17.5 15 6 33% 0.67:1 >40 0.008 0.001 0.0003

Thus, in one embodiment, the thermoreversible sol-gel composition of theinvention comprises:

-   -   a) about 25 to 33% of a mixture of poloxamer 407 and poloxamer        188 in a ratio of between 3:1 and 0.8:1;    -   b) an aqueous salt solution having an ionic strength of 0.05 M        to 0.4 M;    -   c) a phase transition temperature from a liquid phase to a gel        phase of at least 3° C. above or below the temperature of the        surgical field; and    -   d) an elastic modulus of at least 12,000 Pascals in the gel        phase.

In one embodiment, the composition comprises about 25 to 29% of amixture of poloxamer 407 and poloxamer 188 in a ratio of between 3:1 and0.8:1.

In one embodiment, the composition comprises about 27 to 29% of amixture of poloxamer 407 and poloxamer 188 in a ratio of between 1.5:1and 1.6:1, or between 1.53:1 to 1.57:1.

In one embodiment, the composition comprises about 28% of a mixture ofpoloxamer 407 and poloxamer 188 in a ratio of about 1.55:1.

In one embodiment, the salt is selected from the group consisting ofsodium chloride (NaCl), potassium chloride (KCl), sodium sulfate(Na₂SO₄), sodium bisulfate (NaHSO₄), sodium phosphate (Na₃PO₄),monosodium phosphate (NaH₂PO₄), disodium phosphate (Na₂HPO₄), potassiumphosphate (K₃PO₄), monopotassium phosphate (KH₂PO₄), dipotassiumphosphate (K₂HPO₄), various soluble calcium and magnesium salts, such ascalcium chloride (CaCl₂), magnesium chloride (MgCl₂). Other saltsinclude a combination of a cation selected from the group consisting ofsodium, potassium, calcium, magnesium, ammonium, and tetraalkylammonium,with an anion selected from the group consisting of chloride, bromide,tartrate, mesylate, acetate, maleate, and oxalate and otherbiocompatible, water soluble salts include those described in P.Heinrich Stahl, Camille G. Wermuth (Eds.), Handbook of PharmaceuticalSalts Properties, Selection, and Use; 2002. A combination of any two ormore salts can be used.

In one embodiment, the aqueous salt solution has an ionic strength thatis equal to the ionic strength of a solution having a pH of about 7.4and comprising a salt concentration of between 68 mM of sodium chloride,1.3 mM of potassium chloride and 5 mM of phosphate ion to 274 mM ofsodium chloride, 5.4 mM of potassium chloride, and 20 mM of phosphateion.

In one embodiment, the aqueous salt solution is phosphate bufferedsaline.

In one embodiment, the thermoreversible sol-gel composition comprises:

-   -   about 17% w/w of poloxamer 407;    -   about 11% w/w of poloxamer 188;    -   and about 72% of phosphate buffered saline solution.

The compositions of this invention are substantially free frompolyethylene oxide and polyvinylpyrrolidone.

The thermoreversible sol-gel compositions of this invention may includeone or more additional moieties. For example, the composition maycontain a biologically active moiety, preferably in a therapeuticallyeffective amount. Examples of such biologically active moiety includeanti-thrombotic agents, such as anti-coagulants (for example heparin),platelet inhibitors, and thrombolytic agents; anti-anginals, such asbeta-blockers, calcium channel blockers, and nitrates; wound healingagents; growth factors, gene vectors containing growth factors, such asfibroblast growth factor (FGF), insulin-like growth factor (IGF),vascular endothelial growth factor (VEGF), B-cell growth factor (bGF),hepatocyte growth factor (HGF), monocyte chemotactic protein-1 (MCP1),and those described in U.S. Pat. No. 6,702,744, which is incorportatedby reference in its entirety, in protein or gene form, or combinationsthereof, which may be in sustained release formulations; hormones suchas growth hormones, estradiol and other steroids, includingcorticosteroids; pro-angiogenic agents; anti-vessel spasm agents, suchas nitroglycerine; anti-restenosis (anti-proliferative) agents, e.g.,the “limus” family drugs such as everolimus, sirolimus, paclitaxel,rapamycin, biolimus (Biosensors International, Singapore), etc.

In some embodiments, the thermoreversible sol-gel compositions of thisinvention are substantially free of anti-infectives, such asantibiotics, antibacterial, antiviral and antifungal agents;antimicrobials; anti-inflammatory agents, such as diclofenac; andanti-glaucoma agents.

The compositions may also include a contrasting agent. Contrastingagents, such as a biocompatible radio opaque material capable of beingmonitored by, for example, radiography, may also be added to the sol-gelcomposition to track and monitor the sol-gel and/or the procedure. Thecontrasting agent may be water soluble or water insoluble and preferablydoes not contain radioactivity above the native or endogenous amountsnaturally occurring in the elements employed.

Examples of water soluble contrasting agents include metrizamide,iopamidol, iothalamate sodium, iodomide sodium, and meglumine. Examplesof water insoluble contrasting agents include tantalum, tantalum oxide,and barium sulfate, each of which is commercially available in theproper form for in vivo use including a preferred particle size of about10 microns or less. Other water insoluble contrasting agents includegold, tungsten, and platinum powders.

The compositions may also include a suitable biocompatible dye forvisualization, especially when a lumen has a thick wall. Such dyes arewell known in the art.

III. ANASTOMOSIS METHODS

In another aspect, this invention provides methods of using athermoreversible sol-gel composition of this invention to join twonon-conjoined lumens in a patient, which methods comprise:

-   -   a) applying a thermoreversible sol-gel composition of this        invention (as described above) in at least the distal portion of        at least one of the lumens in a manner which imparts structural        integrity to said portion of the lumen or lumens;    -   b) aligning the lumens;    -   c) joining the aligned lumens to form a conduit; and    -   d) removing the composition thereby establishing body fluid flow        through the conduit.

It is contemplated that the present invention can be applied to anyanastomotic procedure that connects one hollow tissue structure (lumen)to another hollow tissue structure (lumen), such that the spaces withineach hollow tissue structure are connected thereby forming a conduit (anintraluminal conduit). It can be used in a microvascular context, whichis performed between ends of blood vessels in the course of, forexample, reattaching severed body part and/or transplanting organ ortissue. It can also be used in minimally invasive procedures orpercutanteous approaches with catheters. The methods of the inventioncan be used to connect non-conjoined lumens arising from surgicalprocedures wherein the originally intact lumen has been severed for thepurposes of, e.g., removing a blockage or partial blockage. Suitablelumens include, by way of example, the vasculature, the vas deferens,the fallopian tubes, the urinary tract, tear ducts, bowel, mammaryglands, alimentary ducts, pancreatic ducts, bile ducts, etc. (Specificanatomical lumens may be referenced by their conventional anatomicalnomenclature such as tubes, ducts or vessels, as used in contextherein.)

In one embodiment, a method of this invention comprises:

-   -   a) applying in a gel phase a thermoreversible sol-gel        composition of this invention (which is described above in        details) into at least the distal portion of at least one of the        lumens;    -   b) aligning the lumens;    -   c) joining the aligned lumens to form a conduit; and    -   d) removing the composition thereby establishing body fluid flow        through the conduit.

In another embodiment, the method comprises:

-   -   a) applying in a solution phase a thermoreversible sol-gel        composition of this invention (which composition is described in        details above) into at least the distal portion of at least one        of the lumens;    -   b) transitioning the sol-gel composition from a liquid phase to        a solid phase;    -   c) aligning the lumens;    -   d) joining the aligned lumens to form a conduit; and    -   e) removing the composition thereby establishing body fluid flow        through the conduit.

In some embodiments, the method is a method for joining at least twonon-conjoined lumens in a patient, wherein at least one of the lumenshas a clamp to arrest the flow of the fluid therein, which methodcomprises:

-   -   (a) providing a biocompatible phase-reversible sol-gel        composition of this invention in a gel phase in at least the        distal portion of at least the lumen having the clamp;    -   (b) aligning the lumens;    -   (c) closing the aligned lumens to form a conduit;    -   (d) inducing a phase transition of the sol-gel wherein the        sol-gel changes from the gel phase to a liquid phase;    -   (e) confirming substantial completion of the phase transition;        and    -   (f) removing the clamp(s) thereby establishing flow through the        conduit and removing the sol-gel composition.

In some embodiments, all non-conjoined lumens have clamps.

In some embodiments, the method is a method for joining at least twonon-conjoined lumens in a patient, which method comprises:

-   -   (a) approximating the lumens in a connecting position;    -   (b) providing a biocompatible phase-reversible sol-gel        composition of this invention in a gel phase in the distal        portion of both lumens;    -   (c) closing the lumens to form a conduit;    -   (d) inducing a phase transition of the sol-gel wherein the        sol-gel changes from the gel phase to a liquid phase; and    -   (e) establishing flow through the conduit.

In some embodiments of the above method, step (e) comprises (e1) and(e2), wherein (e1) comprises confirming substantial completion of thephase transition, and wherein (e2) comprises removing the clamps therebyestablishing flow through the conduit and removing the sol-gelcomposition.

In some embodiments, the approximating the lumens in a connectingposition comprises partially suturing the lumens.

In some embodiments, the above method further comprises a step beforethe step of providing the sol-gel composition in the lumen, which stepcomprises arresting the flow of body fluid in the lumens.

In some embodiments, the arresting the flow of body fluid in the lumencomprises clamping at least one of the lumens. In some embodiments, thearresting the flow of body fluid in the lumen comprises clamping all thelumens to be joined.

In some embodiments, the arresting the flow of body fluid in the lumenscomprises applying a pressure on at least one of the lumens.

In some embodiments, the method further comprises flushing the distalportion of the clamped lumen with a biocompatible liquid prior to thestep of providing the sol-gel composition in the lumen.

In some embodiments, the invention provides a method for joining atleast two non-conjoined lumens in a patient, wherein at least one of thelumens has a clamp to arrest the flow of the fluid therein, which methodcomprises:

-   -   flushing the distal portion of the clamped lumen with a        biocompatible liquid;    -   approximating the lumens in a connecting position, which        approximating comprises partially suturing the lumens;    -   providing a biocompatible phase-reversible sol-gel composition        of this invention in a gel phase in the distal portion of both        lumens;    -   closing the lumens to form a conduit with a biocompatible        adhesive;    -   inducing a phase transition of the sol-gel wherein the sol-gel        changes from the gel phase to a liquid phase;    -   confirming substantial completion of the phase transition; and    -   removing the clamps thereby establishing flow through the        conduit and removing the sol-gel composition.

In one embodiment used for illustrative purposes only, the anastomosisis performed during coronary artery bypass graft (CABG) procedures orperipheral bypass procedures to connect two blood vessels or one bloodvessel with one synthetic graft. The blood vessels connected togethermay have different diameters. Further one or both of the vessels may bevery small, and may be on the order of about 1 to 5 millimeters (“mm”).The microvascular anastomosis procedure using the present invention maybe performed under a microscope.

Referring now to the Figures, the invention is described schematicallyand in a simplistic fashion in order to convey the general concepts.With these concepts in mind those skilled in the art will contemplatedetailed specific embodiments of the invention which are intended to beencompassed by the present claims. FIG. 1 shows a schematiccross-sectional view of a lumen which may be a vessel 1 which has flow 2running there through. A portion of the vessel indicated by points 3 and4 has a restricted flow due to the formation of blockages 5 and 6. Theblockage may become so severe that the flow is completely blocked. Thoseskilled in the art will appreciate that a range of different treatmentsare available for restoring flow.

Prior to the operation to restore flow, the vessel needs to be occludedto stop or reduce the fluid (e.g., blood) flow. Such occlusion isimportant for anastomosis involving a blood vessel to prevent excessiveloss of blood and complications caused by a continuous blood flow. Thiscan be achieved by applying at least one clamp on the blood vessel to beoperated on or by conventional means, such as applying externalpressure, e.g., pressing down with a finger or tournequette, on thevasculature, or providing a vessel loop or vessel tie upstream from thepoint of severance or on both sides of the point of severance on theblood vessel. However, clamping may not be necessary for anastomosis ofother types of lumens where there is no continuous flow of fluid or theamount of fluid does not complicate the procedure.

As shown in FIG. 2, clamps 7 and 8 have been placed on the vessels tostop the blood flow in the section between the clamps prior to removalof the blocked section. Such clamps can be any surgical clamps suitablefor clamping the vessels to be operated on from the outside, such asclamps, clips and tourniquets or snares. After clamping, a portion ofthe vessel 1 which has the restricted flow between the points 3 and 4 isthen surgically removed. Those skilled in the art will understand thatthe distance between the points 3 and 4 is sufficiently small such thatthe ends can be brought into contact with each other to restore flow.

In some embodiments, after the vessel is cut into two non-conjoinedvessels with open ends or after the diseased portion is severed, or inthe case of side-to-end or side-to-side anastomosis after a hole iscreated on the vessel, the part of the vessel between the clamp 7 andthe open end 3 can be flushed with a biocompatible fluid, such as wateror a saline solution, to remove any undesired matter, such as diseasedor dead tissue or cells, mineral deposits, plague, thrombus, bloodclots, etc., that may be present in the vessel. In one embodiment, thesolution is sterile.

Referring now to FIG. 3, the inside of the vessel 1 between the clamp 7and the end 3 has been filled with the thermoreversible sol-gel 10 whichis shown in solid phase by the crossed markings. The area between theclamp 8 and the end 4 is being filled with sol-gel 11 in a liquid phase.In an optional embodiment, the thermoreversible sol-gel 10 can beintroduced in its gel phase so as to ease delivery into the vessel. Thesol-gel is injected from a suitable delivery apparatus such as ahypodermic needle 9. When the sol-gel material is in the hypodermicneedle it may be either in a flowable liquid phase or in a gel phase. Inthe former case, the composition is delivered in a liquid phase at orbelow the ambient temperature of the operating field and will undergo aphase transition to a solid gel phase (10) upon the application of heatto raise the temperature to above the transition temperature. In thelatter case, the delivery apparatus is kept above the transitiontemperature of the thermoreversible sol-gel composition which isdelivered in a solid gel phase (10) and maintained in that phase bymaintaining the temperature at above the transition temperature. It iscontemplated that the later is more convenient when the sol-gelcomposition has a transition temperature of below the temperature of theoperating field. If the sol-gel composition has a transition temperatureof above the temperature of the operating field, the composition can bepreheated and delivered as a gel.

It is contemplated that additional amount of the sol-gel composition maybe applied to the lumens during the joining step if needed.

Alternatively, the procedure includes approximating the lumens in aconnecting position, such as by partially suturing the two lumens. Forexample, a few sutures can be applied on the opposite sides of thelumens to bring the lumens in a connecting position. Thethermoreversible sol-gel composition can then be applied to thepartially sutured opening and form a continuous gel column to fill thelumens to give the structural support for the subsequent procedure thatcompletely joins the lumens. The amount of the sutures applied for thispurpose is not intended to fully close the lumens but is to align thelumens so that the gel can be applied to both lumens together, forexample, in some embodiment, the sutures are 4-6, or 2-4 and preferably1-2 sutures on each side. Approximating the lumens can also be preformedby using temporary or permanent anchoring clamps or clips, laser,electrocautery, etc, that are known in the art to connect lumens.

Alternatively, partial suturing can be applied after application of thegel (making sure there is no twists on the lumen) and before applicationof the glue. For example, the vessels which are deflated due to clampingand lack of blood flow can be inflated with application of a gel. Suturecan then be applied to the inflated vessels to partially connect them tobring them in a connecting position before a glue is applied tocompletely and permanently joining the vessels. The lumens can also bepartially ligated by using temporary or permanent anchoring clamps orclips, laser, electrocautery, etc, that are known in the art forconnecting lumens.

The thermoreversible sol-gel composition is included in a sufficientamount so as to maintain at least a portion of the distal end of vessel1 open. In the absence of some force, the side walls of the vessel 1will contact each other and cause the vessel to close in the absence offlow through the vessel. The solid gel phase (10) imparts sufficientstructural rigidity to the lumen to permit the anastomosis to proceedwith the vessel in its fully filled form.

As shown in FIG. 4, the two lumens can be joined by applying an adhesiveor glue 12 on the two ends of the lumens as well as on the outer surfaceof the vessel at points 3 and 4 and sealed together. Although FIG. 4shows the use of a glue, it is to be understood that as describedherein, joining or ligation can be accomplished any number of ways, suchas suturing or laser soldering or welding. The lumens may also be joinedby using electrocautery or other means known in the art.

Sutures are well known in the art as are surgical glues or adhesives andlasers. It is contemplated that the glues used in the methods of thisinvention are biocompatible and are generally cyanoacrylate-basedadhesives, fibrin-based adhesives, polyurethane-based adhesives,polyisocyanate-based adhesives, polyethylene glycol-based glues, latexglues, biologics glues or protein-based glues, and UV curable glues.Suitable glues include but are not limited to the glues describedherein. The suture or glue may contain biologically active moieties suchas antimicrobial agents (e.g., U.S. Pat. No. 5,762,919).

Laser surgical techniques, such as laser welding or laser soldering, arealso known in the art. See, e.g., D. Simhon, in Lasers in Surgery:Advanced Characterization, Therapeutics, and Systems XIV, Vol. 5312,176-185 (2004); S, Nakata, et al, The Journal of Thoracic andCardiovascular Surgery, 98, 57-62, 1989; and Sanford L. Klein, et al,Microsurgery, 15:4, 287-288, 2005. However, laser anastomoses have beenreported to cause side effects such as soldering the lumen shut,hemorrhage and degeneration of collagen and protein in the adventitiaand media, and under conventional laser anastomosis conditions. See,e.g., S, Nakata, et al, The Journal of Thoracic and CardiovascularSurgery, 98, 57-62, 1989; T. Bavbek, et al, Opthalmologica 219, 267-271,2005. It is contemplated that placing a sol-gel composition inside thelumens will keep the end of the lumen open and protect the tissue frombeing damaged by the laser beam.

Due to the presence of the gel (in solid phase) significant amounts ofadhesive can be used without resulting in vessel closure. In the absenceof a structural support provided by the thermoreversible sol-gel 10within the vessel, the application of pressure to the outside of thevessel, such as the application of the clamps and glue, can causecollapse of the vessel. However, since the thermoreversible sol-gel 10is holding the vessel open, glue can be applied liberally not only tothe ends 3 and 4 which are to be sealed together but the glue 12 can beapplied along the surface of the vessel 1 near the point where the sealis to take place. Thus, as shown in FIG. 4, glue has been applied on theoutside of the vessel 1 on either side of the point where the ends aresealed. The glue can extend outward in any desired amount. However, withsmaller vessels extending the glue out a distance of about 1 mm to about10 mm is generally sufficient. The glue can extend outward around theentire circumference of the connecting point. After the adhesive 12 hasbeen allowed to cure and seal bonds between the two ends of the vessel,the clamps 7 and 8 can be removed.

In one embodiment the glue also penetrates inward to the surface of thegel thereby providing adherence of the entire tissue cross-section ofthe first lumen to the second lumen. In another embodiment, the glue isapplied not only to the outer surface but also to the cross-sectionalsurfaces of the lumens to be closed. It is contemplated that the gluemay be porous to allow tissue ingrowth and healing across the barrier.For example the glue may contain a porogen which may be an emulsifiedliquid that is compatible but is immiscible with the glue and can beabsorbed by the tissue relatively quickly to leave pores in the glue. Itis contemplated that the tissue can infiltrate the glue matrix and healacross as the glue is being reabsorbed by the body. The glue may furthercomprise a biologically active agent as described herein to facilitatetissue growth, prevent or reduce inflection, restenosis, and otherconditions that may be associated with such procedures.

In a still further embodiment, the thermoreversible sol-gel composition,which may have a superior wetting characteristic to the vessel whencompared to the glue, may inhibit the glue from penetrating into theinner surface of lumen. In some embodiments, the glue and the sol-gelhave limited miscibility. When the vessel is not supported by a sol-gel,connecting two vessels with glue may cause safety concerns due to thepossibility that the glue may seal the inside of the lumen. In someembodiments, the glue and the sol-gel are substantially immiscible.

In some embodiments, after the lumens are connected and before theclamps are removed, the gel is changed to its liquid state by, forexample lowering the temperature of the connected lumens to below thetransition temperature of the sol-gel composition by removing the heatsource or by applying a cold source, such as cold water or ice, as shownin FIG. 5A. The operating physician then confirms that the phasetransition is complete, for example by observing the vessel or bywaiting for a sufficient amount of time, to ascertain that no undesiredsolid mass remains in the vessel which may cause safety concerns. Afterconfirming that all sol-gel has become a liquid, the clamps 7 and 8 arethen removed to allow establishment of blood flow and subsequent removalof the liquefied sol-gel by the blood flow. The phase transition can bedone before or after the glue is cured or hardened, preferably after theglue is cured or hardened.

When the clamps 7 and 8 are removed (FIG. 5), the heat which was beingapplied to maintain the composition in a gel state is removed and thegel changes phase to a liquid. Once the gel reverses its phase change tobecome a liquid and the blood flow causes the sol-gel composition to bedispersed, the vessel reopen as shown in FIG. 6. In other cases, thesol-gel composition can be removed by dissolution in the body fluid,such as blood, that will flow to the site of the sol-gel composition. Instill other cases, a combination of phase transition and dissolution canbe used. In some embodiments, the temperature-induced phase transitionand/or dissolution can be facilitated by other means, such as a changein ionic strength, pH, or light, etc.

At this point as shown in FIG. 5, the glue 12 has cured or hardened. Theglue 12 seals the ends of the vessels together but also is applied tothe outer surface of the vessel on either side of the point ofconnection where it is circumferentially applied. Accordingly, when theglue 12 hardens the glue 12 acts as an external stent holding the vessel1 open after the gel has liquefied. Although it is not necessary to havethe glue forming the external stent, this can provide an additionaladvantage. The glue may be designed so that it remains in place for aconsiderable period of time or designed so that it dissolves slowly overtime as the two ends of the vessel grow together.

It is contemplated that by performing an anastomosis procedure using thecompositions and methods of the present invention, the time required toconnect the blood vessels can be significantly reduced compared withusing conventional hand-sewn anastomosis procedures. This in turn canresult in improved outcomes of the procedures in terms of the diametersand patency rates of the vessels connected, anastomotic flow and burststrength, complications caused by the procedures and the mortality ratesof the subjects treated.

The methods of this invention can be applies to end-to-end, end-to-side,and side-to-side anastomosis. It can be used to join two or more bloodvessels in a patient, or join a blood vessel of the a patient with alumen selected from the group consisting of arteriovenous graft,arteriovenous shunt, allograft, xenograft, synthetic graft, and cadaverxenograft.

Other aspects of the above method include visualization or imaging ofthe joining of the lumens and various aspects of procedure as well asmonitoring and evaluating the safety and effectiveness of the procedure.In many cases, the procedure is performed under a condition thatrequires a visual aid in order for the physician to monitor or visualizethe progress of the procedure. For example, a magnifying glass is oftenneeded in an anastomosis procedure. In the microvascular context, anoperating microscope is often used and a laparoscope is used in alaparoscopic procedure. Other imaging medical instruments may also beemployed to monitor the progress and/or the effectiveness of theprocedure, including but not limited to collecting blood samples tomeasure the serum concentration of biomarkers indicative of myocardialperformance or the performance of other organs connected to the lumensbeing conjoined, such as measuring troponin T levels in a coronaryartery bypass grafting procedure, measuring the diameters of theconjoined lumens using, for example CT angiography, determining patencyand flow through, for example ultrasonic Doppler imaging (e.g., using avisual sonic machine), and/or monitoring the presence of the liquefiedsol-gel in the serum or urine. Suitable methods are generally known inthe art.

Thus in another aspect, this invention provides methods for imaging thejoinder of at least two non-conjoined lumens using the methods of thisinvention and the stabilization of the geometry of the distal portionsof the lumens in a patient.

In some embodiments, the method is a method for imaging the joinder(ligation) of at least two non-conjoined lumens and the stabilization ofthe geometry of the distal portion of the lumens in a patient, themethod comprises:

-   -   (a) acquiring an image of a sol-gel composition of this        invention in at least the distal portion of at least one of the        lumens;    -   (b) acquiring an image showing alignment of the lumens and        closure of the aligned lumens to form a conduit.

In some embodiments, the method is a method for imaging the joinder(ligation) of at least two non-conjoined lumens and the stabilization ofthe geometry of the distal portion of the lumens in a patient, whereinat least one of the lumens is clamped, the method comprises

-   -   (a) acquiring an image of a sol-gel composition of this        invention in a gel state in at least the distal portion of at        least one of the lumens in proper position to impart structural        integrity for the non-conjoined lumens;    -   (b) acquiring an image of the proper alignment of the lumens to        be joined;    -   (c) acquiring an image of the closing of the lumens; and    -   (d) acquiring an image of the removal of the clamps.

In some embodiments, the method further comprises acquiring an image ofthe internal walls of the distal portion of the lumen before and/orafter the lumen is flushed with a biocompatible liquid.

In some embodiments, the method is a method for imaging the joining ofat least two non-conjoined lumens and the stabilization of the geometryof the distal portions of the lumens in a patient, the method comprises

-   -   (a) acquiring an image of the approximation of the lumens for a        connecting position;    -   (b) acquiring an image of a biocompatible phase-reversible        sol-gel composition of this invention in a gel phase in the        distal portion of both lumens in proper position for joining the        non-conjoined lumens; and    -   (c) acquiring an image of the closure of the lumens to form a        conduit.

In some embodiments, the method is a method for imaging the joining atleast two non-conjoined lumens and the stabilization of the geometry ofthe distal portions of the lumens in a patient, wherein at least one ofthe lumens has a clamp, the method comprises

-   -   (a) acquiring an image of the internal walls of the distal        portion of the lumen after the lumen is flushed with a        biocompatible liquid;    -   (b) acquiring an image of the approximation of the lumens for a        connecting position;    -   (c) acquiring an image of a biocompatible phase-reversible        sol-gel composition in a gel phase in the distal portion of both        lumens in proper position for joining the non-conjoined lumens;    -   (d) acquiring an image of the closure of the lumens to form a        conduit; and    -   (e) acquiring an image of the removal of the clamps and        establishment of the body fluid.

In another aspect, this invention provides methods for visualizingguidance for the methods of this invention for joining at least twonon-conjoined lumens in a patient.

In some embodiments, the method is a method for visualizing guidance forjoining at least two non-conjoined lumens and the stabilization of thegeometry of the distal portions of the lumens in a patient, the methodcomprises

-   -   (a) displaying the provision of a sol-gel composition of this        invention in a gel phase in at least the distal portion of at        least one of the lumens in proper position to impart structural        support for the non-conjoined lumens;    -   (b) visualizing the proper alignment of the lumens to be joined;    -   (c) visualizing the closing of the lumens form a conduit; and    -   (d) visualizing the induction of the phase change of the sol-gel        composition in the lumen where the sol-gel changes from the gel        phase to a flowable liquid phase.

In some embodiments, the method is a method for visualizing guidance forjoining at least two non-conjoined lumens and the stabilization of thegeometry of the distal portions of the lumens in a patient, wherein atleast one of the lumens has a clamp, the method comprises

-   -   (a) displaying the provision of a sol-gel composition of this        invention in at least the distal portion of at least one of the        lumens in proper position to impart structural support for the        non-conjoined lumens;    -   (b) visualizing the proper alignment of the lumens to be joined;    -   (c) visualizing the closing of the lumens to form a conduit; and    -   (e) visualizing the removal of the clamps.

A method for visualizing guidance for joining at least two non-conjoinedlumens and the stabilization of the geometry of the distal portions ofthe lumens in a patient, the method comprises

-   -   (a) visualizing the approximation of the lumens for a connecting        position;    -   (b) displaying the provision of a biocompatible phase-reversible        sol-gel composition in its gel phase in the distal portion of        both lumens in proper position for joining the non-conjoined        lumens; and    -   (c) monitoring by visualization closure of the lumens to form a        conduit.

In some embodiments, the method further comprises visualizing theinternal walls of the distal portion of the lumen after the lumen isflushed with a biocompatible liquid.

In some embodiments, the method further comprises visualizing the flowof body fluid through the conduit.

A method for visualizing guidance for joining at least two non-conjoinedlumens and the stabilization of the geometry of the distal portions ofthe lumens in a patient, wherein at least one of the lumens has a clamp,the method comprises

-   -   (a) visualizing the internal walls of the distal portion of the        lumen after the lumen is flushed with a biocompatible liquid;    -   (b) visualizing the approximation of the lumens for a connecting        position;    -   (c) displaying the provision of a biocompatible phase-reversible        sol-gel composition in a gel phase in the distal portion of both        lumens in proper position for joining the non-conjoined lumens;    -   (d) monitoring by visualization closure of the lumens to form a        conduit; and    -   (e) visualizing the flow of body fluid through the conduit.

In another aspect, this invention provides a method for evaluating theeffectiveness of the methods of this invention for joining non-conjoinedlumens in a patient.

In some embodiments, the method is a method for evaluating theeffectiveness of surgical joining of at least two non-conjoined lumensin a patient using a thermoreversible gel, the surgical joiningprocedure comprising a clamp on at least on one of the lumens to arrestthe flow of the fluid therein and the thermoreversible gel in at leastthe distal portion of at least one of the lumens, alignment of thelumens, closure of the aligned lumens to form a conduit, induction of aphase change of the gel to a flowable liquid, substantial completion ofthe phase change and removal of the clamp and flowable liquid, theevaluation method comprising

-   -   (a) measuring the serum concentration of biomarkers indicative        of myocardial performance or the performance of other organs        connected to the conjoined lumens; and/or    -   (b) measuring the diameters of the conjoined lumens, for example        using CT angiography; and/or    -   (c) determining patency and flow, for example through ultrasonic        Doppler imaging; and/or    -   (d) monitoring the presence of the flowable liquid in the serum        or urine.

It is contemplated that the compositions and methods of the presentinvention are useful in other medical procedures, such as reversal ofvasectomy, reversal of fallopian tube ligation, and reconstructive tubalsurgeries to treat blocked or damaged fallopian tubes. The method canalso be used to connect an AV graft or AV shunt or fistula to a bloodvessel for hemodialysis as well as below the knee bypass, such as in thetreatment of a peripheral arterial disease. Further the compositions andmethods of the present invention can be used in alimentary anastomosis.Significant leak rates (about 2-5%) have been resulted by currentalimentary anastomosis procedures. Many of the leak incidents are fatalor lead to significant morbidity. Because the alimentary tubes beingligated can be supported inside by the composition of the presentinvention and the glue can be applied circumferentially outside thepoint of connection, thus allowing complete sealing, it is contemplatedthat anastomosis using the compositions and methods of the presentinvention will significantly reduce the leak rate and lead to decreasedmortality and morbidity caused by alimentary anastomosis. The methodsand compositions of the present invention can also be used in thetreatment of conditions involving urinary tracts, tear ducts, bowel,mammary ducts, pancreatic ducts, bile ducts, and the like.

IV. KITS OF THE INVENTION

One aspect of this invention is in the form of a kit of parts. The kitmay include specific instructions with respect to how to carry out themethodology of the invention as exemplified above.

Further, the kit may include a container containing the thermoreversiblesol-gel composition of the invention, preferably in sterile form, and adelivery device. The delivery device may be a syringe, a pipette,catheter or laparoscopic tool, or the like. Alternatively, the kit maycontain a delivery device loaded with the thermoreversible sol-gelcomposition, again preferably in sterile form. For example, the kit maycontain a syringe already loaded with a sterile sol-gel composition oran ampule made from glass or plastic that contains a sterile sol-gelcomposition, and which has a tip that may be cut open to apply thesol-gel contained therein.

Still further, the kit may include one, two or more clamps of the typewhich might generally be used in connection with the lumen or type ofvessel (or duct, tube, etc.) being treated. Still further, the kit mayinclude sutures, anchoring clamps or clips or surgical glue of the typedescribed above. Still further, the kit may include a heat source toinduce a liquid to gel transition and to maintain the sol-gel in itssolid or gel like phase until it is to be removed. Such heat source maybe a heated air blower capable of delivering warm air, heating pads, alight energy source, an irrigated liquid stream source, an infraredheating apparatus or the like, that can maintain a temperaturesufficient to induce the change of phase of the thermoreversiblesol-gel. Further, the delivery device may be coupled with a heat orcooling component and serve as a heating or cooling source. The kit mayalso include a cooling source to induce a gel to liquid transition whenit is to be removed. The kit may further include an adhesive deliverysystem. The kit may further include one or more pharmaceutically activedrugs which may be separate from or incorporated into thethermoreversbile sol-gel composition of this invention. Thus, forexample, the drug which may be provided separately in the kit orincorporated into the thermoreversible sol-gel composition may includean anticoagulant such as heparin. The thermoreversible sol-gelcomposition may further include other materials such as a wound healingmedicament which aids in healing of the vessel.

Commercially, for ease in practical application, materials may beprepared so that they are sterile and substantially pyrogen free, forexample, in accordance with regulatory requirements. Materials anddevices may be prepackaged in sterile packaging.

V. EXAMPLES Example 1 Preparation and Property Determination of aSol-Gel Composition of this Invention

Presented here is a protocol for preparing 100 mL (100 g) of athermoreversible sol-gel composition having Formulation 1 listed inTable 1.

Materials:

-   -   Phosphate buffered saline (“PBS”): 72 g, pH 7.4, with Ca²⁺ and        Mg²⁺;    -   Poloxamer 407 (BASF Pluronic® F 127): 17 g;    -   Poloxamer 188 (BASF Pluronic® F 68): 11 g.

Procedure:

The above materials were mixed at 4° C. under high shear to facilitatedispersing the solid poloxamer in the solvent. The heterogenous mixtureis then incubated at 4° C. for 12 hours to allow for completedissolution of the poloxamer in the solvent. The mixture is then furtherstirred to ensure homogeneity.

Compositions having other formulations can be made similarly accordingto the total concentration of the poloxamers and the ratios of the twopoloxamers.

Water or other aqueous solutions may replace PBS as the solvent. It ispreferred that the solvent has an ionic concentration and pH that areclose to that of the body fluid so that the resulting thermoreversiblesol-gel compositions do not cause substantial damage to surroundingtissues.

To determine the sol-gel transition temperature (the temperature atwhich the composition transitions from liquid to gel), amounts of eachsolution were placed between the parallel plates of a stress rheometer(a TA Instruments AR1500EX rheometer was used). The plates were 4 cm indiameter and the gap was set to 1 mm. The elastic modulus was measuredusing the rheometer at a frequency of 1 Hz and a strain of 1% atdifferent temperatures. Table 1 shows the modulus of several differentformulations of the sol-gel composition at various temperatures.

Example 2 An Anastomosis Operation Using a Sol-Gel Composition of thisInvention

End to End Anastomosis Surgical Method Performed on Cardiac TubularTissues in Rats:

Rats, which are treated and cared for in accordance with all applicablelaws and regulations, and in accordance with good laboratory practices,are anesthetized with isoflurane and prepped in sterile fashion with 70%ethanol. The aorta is exposed and isolated with blunt and sharpdissection through a midline laparotomy incision. The aorta is clampedproximally and distally, and subsequently divided and flushed withheparinized saline. In some cases the rat and a sol-gel composition ofFormulation 1 are warmed to about 37° C. with sterile water, and thenthe sol-gel composition of Formulation 1 is injected into severed aortasin a semi-solid state. In other cases the sol-gel composition ofFormulation 1 is injected as a liquid and then heated to about 37° C.with a convectional source to solidify it. After direct reapproximationby pushing the ends of the aorta together, the cyanoacrylate adhesive isapplied and allowed to cure for 60-120 seconds. The clamps are thenremoved, and the midline incision is closed in layers with 5-0 vicryl(Ethicon, Inc., Somerset, N.J.) and 4-0 nylon (Ethicon) in a runningfashion.

End to Side Anastomosis Surgical Method:

Rats are anesthetized, shaved, and prepped in standard sterile fashion.A midline laparotomy incision is made and the abdominal organs areeviscerated into a moist 4×4 gauze. The iliac bifurcation is isolatedwith blunt dissection, and the abdominal aorta is clamped proximally andthe iliacs are clamped distally. The left iliac is divided just distalto the bifurcation and an arteriotomy is made in the right iliac. Thevessels are then flushed with heparinized saline. The abdomen is warmedat the same time a sol-gel composition of Formulation 1 is warmed to agel (about 37° C.) which is then injected. The left iliac isapproximated to the right iliac using the sliding clamps (one clamp ison the proximal aorta and the other is on the distal left iliac). Oncethe ends are opposed without tension, the cyanoacrylate is applied andallowed to set (˜5 min). The clamps are then removed and blood flow isrestored.

Example 3 Prophetic Example of Using a Sol-Gel Composition in a VascularGrafting Procedure

In clinical practice, there are some instances where native vessels(i.e. blood vessels of the patient) are not available for bypassgrafting or where, especially for older patients, native fistulasrequire a long maturation time or do not develop sufficiently. In theseinstances, physicians may choose to use artificial conduits such asgrafts comprising polytetrafluoroethylene (PTFE), which can be formed inreinforced or nonreinforced configurations, for example expended PTFE(ePTFE) grafts, manufactured by Gore and others. Synthetic internalarteriouvenous (AV) fistulas or AV grafts, can be placed in manypositions in the arms or legs or across the anterior chest wall.

The artificial grafts may be used instead of native vessels in allconfigurations: end-to-end inter-positional grafting, end-to-side forprocedures such as artery to vein shunt for dialysis fistula access,end-to-side bypass procedure both for coronary bypass as well asperipheral bypass. For example, in the below the knee bypass, the graftis attached proximally to the femoral artery and then distally to thepopliteal artry and this conduit carries the blood to the distal limb.The procedure for using the graft is similar to the ones for nativevessels described above, where the ends of the non-conjoined lumen(expect here one native and one graft) are filled with the gel, followedby alignment of the ends, depositing of glue over the joint, dissolutionof the gel is dissolved to complete the anastomosis.

The preceding merely illustrates the principles of the invention. Itwill be appreciated that those skilled in the art will be able to devisevarious arrangements which, although not explicitly described or shownherein, embody the principles of the invention and are included withinits spirit and scope. Furthermore, all examples and conditional languagerecited herein are principally intended to aid the reader inunderstanding the principles of the invention and the conceptscontributed by the inventors to furthering the art, and are to beconstrued as being without limitation to such specifically recitedexamples and conditions. Moreover, all statements herein recitingprinciples, aspects, and embodiments of the invention as well asspecific examples thereof, are intended to encompass both structural andfunctional equivalents thereof. Additionally, it is intended that suchequivalents include both currently known equivalents and equivalentsdeveloped in the future, i.e., any elements developed that perform thesame function, regardless of structure. The scope of the presentinvention, therefore, is not intended to be limited to the exemplaryembodiments shown and described herein. Rather, the scope and spirit ofpresent invention is embodied by the appended claims.

1. A thermoreversible sol-gel composition for surgical use having aphase transition temperature from a liquid phase to a gel phase of atleast 1° C. above or below the temperature of the surgical field and agel phase elastic modulus of at least 8,000 Pascals, which compositioncomprises at least two poloxamers in an aqueous solvent.
 2. Thecomposition of claim 1, wherein the transition temperature is at least3° C. above or below the temperature of the surgical field. 3.(canceled)
 4. The composition of claim 1, which composition is sterile.5. The composition of claim 1, wherein the phase transition temperatureis about 33 to 38° C.
 6. (canceled)
 7. The composition of claim 1,wherein the elastic modulus of the gel phase is at least 10,000 Pascals.8. (canceled)
 9. The composition of claim 1, which composition has a pHof about 6 to
 8. 10. The composition of claim 1, wherein the aqueoussolvent is water, or a water-based solution with or without salt. 11.The composition of claim 1, comprising: a) about 25 to 33% of a mixtureof poloxamer 407 and poloxamer 188 in a ratio of between 3:1 and 0.8:1;and b) an aqueous salt solution having an ionic strength of from about0.05 M to about 0.4 M.
 12. The composition of claim 11, comprising about27 to 29% of a mixture of poloxamer 407 and poloxamer 188 in a ratio ofbetween 1.5:1 and 1.6:1. 13-14. (canceled)
 15. The composition of claim11, wherein the salt is selected from the group consisting of sodiumchloride, potassium chloride, sodium sulfate, sodium bisulfate, sodiumphosphate, monosodium phosphate, disodium phosphate, potassiumphosphate, monopotassium phosphate, dipotassium phosphate, calciumchloride, magnesium chloride, and a combination thereof. 16-17.(canceled)
 18. The composition of claim 17, wherein the aqueous saltsolution is phosphate buffered saline.
 19. The composition of claim 1,comprising about 17% w/w of poloxamer 407; about 11% w/w of poloxamer188; and about 72% of phosphate buffered saline solution.
 20. Thecomposition of claim 1, further comprises a biologically active agent, abiocompatible dye, or a biocompatible contrasting agent or combinationsthereof.
 21. The composition of claim 20, wherein the biologicallyactive agent is selected from the group consisting of an anti-coagulant,anti-spasm agents, anti-restenosis agents, pro-angiogenic agents,fibroblast growth factor, insulin-like growth factor, vascularendothelial growth factor, hepatocyte growth factor, monocytechemotactic protein-1, nitroglycerine, everolimus, sirolimus,paclitaxel, rapamycin, biolimus, and combinations thereof, and furtherwherein the biologically active agent is optionally formulated insustained release or in gene vectors.
 22. The composition of claim 21,wherein the biologically active agent is heparin. 23-26. (canceled) 27.A method for joining at least two non-conjoined lumens in a patientwhich method comprises: (a) applying a composition of claim 1 in atleast the distal portion of at least one of the lumens in a manner whichimparts structural integrity to said portion of the lumen; (b) aligningthe lumens; (c) joining the aligned lumens to form a conduit; and (d)removing the composition thereby establishing body fluid flow throughthe conduit.
 28. A method for joining at least two non-conjoined lumensin a patient, wherein at least one of the lumens has a clamp to arrestthe flow of the fluid therein, which method comprises: (a) providing abiocompatible phase-reversible sol-gel composition of claim 1 in a gelphase in at least the distal portion of at least one of the lumens; (b)aligning the lumens; (c) closing the aligned lumens to form a conduit;(d) inducing a phase transition of the sol-gel wherein the sol-gelchanges from the gel phase to a liquid phase; (e) confirming substantialcompletion of the phase transition; and (f) removing the clamp(s)thereby establishing flow through the conduit.
 29. A method for joiningat least two non-conjoined lumens in a patient, which method comprises:(a) approximating the lumens in a connecting position; (b) providing abiocompatible phase-reversible sol-gel composition of claim 1 in a gelphase in the distal portion of both lumens; (c) closing the lumens toform a conduit; (d) inducing a phase transition of the sol-gel whereinthe sol-gel changes from the gel phase to a liquid phase; and (e)establishing flow through the conduit. 30-35. (canceled)
 36. A methodfor joining at least two non-conjoined lumens in a patient, wherein atleast one of the lumens is clamped to arrest the flow of body fluidtherein, which method comprises: flushing the distal potion of theclamped lumen with a biocompatible liquid; approximating the lumens in aconnecting position, which approximating comprises partially suturingthe lumens; providing a biocompatible phase-reversible sol-gelcomposition of claim 1 in a gel phase in the distal portion of bothlumens; closing the lumens to form a conduit with a biocompatibleadhesive; inducing a phase transition of the sol-gel wherein the sol-gelchanges from the gel phase to a liquid phase; confirming substantialcompletion of the phase transition; and removing the clamps therebyestablishing flow through the conduit and removing the sol-gelcomposition. 37-45. (canceled)
 46. The method of claim 27, wherein thelumens are joined by applying a biocompatible adhesive between an end ofthe first lumen and an end of the second lumen and allowing the adhesiveto form bonds between the lumens.
 47. The method of claim 46, whereinthe biocompatible adhesive is selected from among a cyanoacrylate-basedadhesive, a fibrin-based adhesive, a polyurethane-based adhesive, alatex glue, a biologics glue, a polyethylene glycol-based glue, and apolyisocyanate-based adhesive.
 48. The method of claim 46, wherein theadhesive is applied to the cross-sectional surfaces of the lumens. 49.The method of claim 46, wherein the adhesive is applied around thecircumference of the joined lumens.
 50. The method of claim 46, whereinthe adhesive further forms an external stent.
 51. The method of claim27, wherein the lumens are joined by a method comprising sutures,electrocautery, or applying laser.
 52. The method of claim 27, furthercomprising an extra step of partially suturing the lumens, where theextra step is performed before and/or after step (a).
 53. (canceled) 54.The method of claim 27 or 36, wherein a first lumen is a vessel of thecardiovascular system of a human.
 55. The method of claim 54, wherein asecond lumen is selected from the group consisting of a vessel of thecardiovascular system of a human, an arteriovenous graft, an allograft,a xenograft, cadaver xenograft, a synthetic graft, and an arteriovenousshunt. 56-57. (canceled)
 58. The method of claim 27 or 36, wherein atleast one of the lumens has a diameter of less than 1 mm.
 59. The methodof claim 27 or 36, wherein the at least two lumens are selected from thegroup consisting of human fallopian tubes, vas deferens, tubes in thealimentary canal, pancreatic ducts, bile ducts, tear ducts, and mammaryducts.
 60. The method of claim 27 or 36, wherein the lumens are joinedend to end, end to side or side to side. 61-62. (canceled)
 63. A vesselof a cardiovascular system of a mammal, wherein the vessel has acomposition of claim 1 positioned in the vessel in a manner and in anamount sufficient to hold walls of the vessel open in a fashion similarto when blood flows through the vessel.
 64. A kit for connecting atleast two non-conjoined lumens within a living mammal, comprising: adelivering device; and a composition of claim
 1. 65. The kit of claim64, wherein the delivering device is a syringe.
 66. The kit of claim 65,wherein the composition is loaded in the syringe.
 67. The kit of claim64, where delivery device is a catheter or a laparoscopic tool.
 68. Thekit of claim 64, wherein the delivery device is coupled with a heatingor cooling component.
 69. The kit of claim 64, further comprising atleast one clamp for arresting the flow of body fluid in at least one ofthe non-conjoined lumen, a biocompatible adhesive for sealing the lumensor a heat source.
 70. (canceled)
 71. A method for imaging the geometryof the distal portion of at least one lumen being supported by a sol-gelcomposition of claim 1 and the joinder of the lumen with at leastanother lumen in a patient.
 72. A method for visualizing guidance of thegeometry of the distal portion of at least one lumen being supported bya sol-gel composition of claim 1 and the joinder of the lumen with atleast another lumen in a patient.
 73. A method for evaluating theeffectiveness of a method for joining at least two non-conjoined lumensin a patient wherein at least one of the non-conjoined lumens having asol-gel composition of claim 1 to provide geometry support for thelumen.